Toxicological evaluation of preservative-containing and preservative-free topical prostaglandin analogues on a three-dimensional-reconstituted corneal epithelium system
- Hong Liang1,2,3,4,5,
- Aude Pauly2,3,4,5,
- Luisa Riancho2,3,4,5,
- Christophe Baudouin1,2,3,4,5,6,
- Françoise Brignole-Baudouin2,3,4,5,7
- 1Department of Ophthalmology III, Quinze-Vingts National Ophthalmology Hospital, Paris, France
- 2Institut National de la Santé et de la Recherche Médicale, U968, Paris, France
- 3Université Pierre et Marie Curie University Paris, France
- 4Unité Mixte de Recherche en Santé 968, Institut de la Vision, Paris, France
- 5Centre National de la Recherche Scientifique, Unité Mixte de Recherche 7210, Paris, France
- 6Ambroise Paré Hospital, APHP, University of Versailles Saint-Quentin-en-Yvelines, Versailles, France
- 7Department of Toxicology, Faculty of Biological and Pharmacological Sciences, Paris, France
- Correspondence to Dr Françoise Brignole-Baudouin, UMR_S968, INSERM, Institut de la Vision, 17 Rue Moreau, Paris 75012, France;
- Accepted 2 January 2011
- Published Online First 22 March 2011
Aims Using an established three-dimensional (3D) toxicological model based on reconstituted human corneal epithelium (HCE), this study investigated the tolerability of four topical intraocular-pressure-lowering agents: the commercial solutions of benzalkonium chloride (BAC)-containing 0.005% latanoprost, 0.004% travoprost, 0.03% bimatoprost containing 0.02%, 0.015% and 0.005% BAC, respectively, and the preservative-free (PF) tafluprost. Solutions of 0.01% and 0.02% BAC alone were also evaluated for comparison.
Methods The 3D-HCEs were treated with solutions for 24 h followed or not by a 24 h recovery period. We used a modified MTT (3(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) procedure to assess cell viability in the HCE. Frozen sections of HCE were analysed using fluorescence microscopy for the evaluation of apoptosis (terminal deoxynucleotidyl transferase mediated dUTP nick end labelling), inflammation (ICAM-1) and proliferation (Ki67). Corneal epithelial tight junctions (occludin and tight junction protein 1 (zona occludens 1)) were also assessed by en face confocal microscopy in response to the different eye-drops.
Results The MTT test revealed that the cytotoxicity of antiglaucoma eye-drops was primarily related to the concentration of their common BAC preservative (0.02% BAC-latanoprost>0.015% BAC-travoprost>0.005% BAC-bimatoprost). PF-tafluprost did not induce any obvious cytotoxicity, showed the least expression of inflammatory or apoptotic markers and revealed preservation of membrane immunostaining of tight junction proteins in comparison with BAC-containing solutions.
Conclusion The toxicological model of the 3D reconstructed corneal epithelia model confirmed the ocular surface cytotoxicity of BAC-containing antiglaucomatous solutions. Compared with the formulations containing the toxic preservative BAC, PF-tafluprost was well tolerated without inducing significant corneal epithelium deterioration.
Funding The study was supported by an unrestricted grant from Santen Oy (Tempere, Finland).
Competing interests None.
Provenance and peer review Not commissioned; externally peer reviewed.
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